Studies on the mechanisms and control of energy-transducing systems in normal and tumor cells will be continued. Three ATP-driven ion pumps from normal tissues will be analyzed: the H+pump of clathrin-coated vesicles from bovine brain, the Ca2+pump of sarcomplasmic reticulum of rabbit muscle, and the Na+K+ATPase from dog kidney. The proton pump has been solubilized and will be purified to homogeneity. A hydrophobic membrane fragment of the clathrin-coated vesicle was shown to be required for reconstitution. This will also be purified. The Ca2+pump will be reconstituted with synthetic and natural phospholipids and proteolipids, and conditions regulating the efficiency of pumping will be explored. Similar studies will be performed with the Na+K+ATPase. Glycolysis in tumor cells will be analyzed with respect to the ATPases required for the delivery of ADP and Pi. We shall explore how oncogenes increase the rate of aerobic glycolysis. Growth factors such as TGFs known to stimulate glycolysis will be analyzed and their connections with oncogenes explored. A recent discovery showing that methionine at 10 mM inhibits glycolysis of transformed cells or untransformed cells that have been exposed to transforming factor beta will be further analyzed in great detail. (E)